Technical Data Sheet
Readycult®
Coliforms 100
Ordering number: 1.01298.0001
Selective enrichment broth for the simultaneous presence / absence detection and identification of total
coliforms and E. coli within the bacteriological water examination.
Readycult® Coliforms 100 Presence/Absence Test for Detection and Identification of Coliform Bacteria
and Escherichia coli in Finished Waters has been reviewed and approved by USEPA’s Office of Ground
Water and Drinking Water as acceptable method for monitoring total coliforms and E. coli under the
Total Coliform Rule (40 CFR 141.21). This version incorporated changes that were approved in May
2005 (ATP Case No. BD03-001).
General Information
Content: 20 snap packs
1 snap pack each for 100 ml of water sample.
Mode of Action
The high nutritional quality of the peptones and the incorporated phosphate buffer guarantee rapid
growth of coliforms whereas lauryl sulfate largely inhibits the accompanying flora, especially the Gram-
positive. By adding the chromogenic substrate X-GAL which is cleaved by coliforms and the fluorogenic
substrate MUG which is highly specific for E. coli the simultaneous detection of total coliforms and E.
coli is possible. The presence of total coliforms is indicated by a blue-green color of the broth and E. coli
by a blue fluorescence under UV-light.
The life science business of Merck operates as
MilliporeSigma in the U.S. and Canada. 1 of 4
Typical Composition (g/L)
Readycult® Coliforms 100
Tryptose 0.5
Sodium chloride 0.5
Sorbitol 0.1
Tryptophan 0.1
di-Potassium hydrogen phosphate 0.27
Potassium dihydrogen phosphate 0.2
Lauryl sulfate sodium salt 0.01
X-GAL 0.008
MUG 0.005
IPTG 0.01
pH: 6.8 ± 0.2 at 25°C
Preparation
1. Add 100 ml of water sample into a sterile, transparent vessel with screw cap. (minimum capacity:
>120 ml) Attention: please use material e.g. glass that is not self-fluorescing!
2. Take one snap pack, shortly tap to ensure the granules are at the bottom. Bend the upper part of the
snap pack until it breaks open. Attention: do not touch the opening to avoid contamination risk!
3. Add the content to the water sample. Seal the vessel and shake to dissolve the granules completely.
The prepared broth is clear and yellowish.
Incubation: up to 24 h at 35°C to 37°C. If incubated at room temperature (20 to 25°C) the incubation
time is prolonged to 48 hours
Experimental Procedure and Evaluation
Interpretation of results for the detection of Total Coliforms / E. coli:
Negative: No color change.
The broth remains yellowish in color.
Total coliforms: Any color change of the broth to blue-green, even in the upper section of the broth
only, confirms the presence of coliforms (X-GAL reaction). No decoloration with shaking!
E. coli: Check blue-green colored vessels for fluorescence by using a long wave UV-lamp (366 mm) in
front of the vessel. Light blue fluorescence confirms the presence of E. coli (MUG reaction). Further
confirmation or verification steps are not required.
Attention: Protect your eyes from direct UV light!
We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowledge and 2 of 4
ability, but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies
in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking
the suitability of our products for the envisaged purpose.
Merck, GranuCult, Readytube and Readyplate are trademarks of Merck KGaA, Darmstadt, Germany. Anaeroclip, Anaerocult, Bactident,
ChromoCult, Duopath, FluoroCult, Readycult, Readybag and Singlepath are registered trademarks of Merck KGaA, Darmstadt,
Germany.
Optional: Further verification of E. coli in the vessel with positive fluorescence is possible with Raedycult
Coliforms: Overlay the broth with 2.5 ml of KOVAC's reagent (indole reaction). A red ring confirms
presence of E. coli. This step is not mandatory for the confirmation of E.coli !
Colour change to Indole-
Fluorescence
blue-green Reaction (optional)
Total coliforms + - -
E. coli + + +
yellow colour
Negative
(no change)
Disposal
Autoclave the broth (15 min/121°C). Alternatively heat the broth for 30 min. in boiling water or use a
proper disinfectant.
Storage
In case the sample is to be stored below 25°C, the examination has to be started within 6 hours.
Exceptionally the sample can be stored at 2 to 8°C (refrigeration) for up to 24 hours.
Store dry at 15°C to 25°C.
If stored under recommended conditions the unopened snap pack has a shelf-life of 3 years after day
of production (see expiry date on the label).
Quality Control
Inoculum
Color
on
Control strains Growth change to Fluorescence Indole formation
reference
blue-green
medium
Escherichia coli
ATCC 11775 10-100 + + + +
(WDCM 00090)
Citrobacter
freundii 10-100 + + - -
ATCC 8090
Salmonella
typhimurium
10-100 + - -
ATCC 14028 -
(WDCM 00031)
Klebsiella
pneumoniae 10-100 + + - -
ATCC 31488
Pseudomonas
aeruginosa
≥1000 No limit - - -
ATCC 10145
(WDCM 00024)
Incubation: 24 h; 35°C; aerobic
Please refer to the actual batch related Certificate of Analysis.
We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowledge and 3 of 4
ability, but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies
in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking
the suitability of our products for the envisaged purpose.
Merck, GranuCult, Readytube and Readyplate are trademarks of Merck KGaA, Darmstadt, Germany. Anaeroclip, Anaerocult, Bactident,
ChromoCult, Duopath, FluoroCult, Readycult, Readybag and Singlepath are registered trademarks of Merck KGaA, Darmstadt,
Germany.
Literature
USEPA United States Environmental Protection Agency – National Primary Drinking Water Regulations
- Analytical Methods Approved for Drinking Water Compliance Monitoring under the Total Coliform Rule.
Revised December 2009
Ordering Information
Product Cat. No. Pack size
Readycult® Coliforms 100 1.01298.0001 1 x 20 tests
Bactident® Indole (dropper bottle) 1.11350.0001 1 x 30 ml
CULTURA® Mini-Incubator (100-110 V) 1.15533.0001 1 ea
CULTURA® Mini-Incubator (220-235 V) 1.13311.0001 1 ea
KOVCS Indole Reagent 1.09293.0100 100 ml
UV Lamp (366 nm) 1.13203.0001 1 ea
Merck KGaA Find contact information for your
country at:
Frankfurter Strasse 250
www.merckmillipore.com/offices
64293 Darmstadt, Germany
Fax: +49 (0) 61 51 / 72-60 80 For Technical Service, please visit:
www.merckmillipore.com/techservice
For more information, visit
www.merckmillipore.com/biomonitoring
We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowledge and 4 of 4
ability, but without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies
in respect to any rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking
the suitability of our products for the envisaged purpose.
Merck, GranuCult, Readytube and Readyplate are trademarks of Merck KGaA, Darmstadt, Germany. Anaeroclip, Anaerocult, Bactident,
ChromoCult, Duopath, FluoroCult, Readycult, Readybag and Singlepath are registered trademarks of Merck KGaA, Darmstadt,
Germany.
